Abstract: Phosphofructokinase(PFK)is one of the important regulatory enzymes in the Embden Meyerhof Parnas pathway.The PFK gene(pfkA)was cloned from the wine-brewing Aspergillus oryzae,CICC2012.It was subsequently sequenced and submitted to Genbank(accession number:KC113503.1)for verification.The sequence analysis revealed that pfkA was 2,722bp in length containing two introns,and 2,358bp long with an open reading frame encoding 785amino acids.The PFK was a hydrophilic protein containing two family signatures with a molecular weight of 86.0kD and an isoelectric point of 6.38.Its secondary structure consisted of 42.68% α-helix,14.27%β-folding and 43.06% random coil.The 3Dhomology model showed two(i.e.,N-and C-terminal) domains with the substrate,fructose-6-phosphate,bound to the N-terminal domain.The phylogenetic tree indicated pfkA′s close relationship with PFKs from other filamentous fungi.